Dear Rita-

Your problem sounds more like a variation in section thickness than a
staining phenomenon.  If one whole section out of six is light (assuming
this lightness is consistent over the section), it is very hard indeed to
think of a way the staining could be the culprit.  That is, of course,
assuming the light section is in the middle of the slide, not the very top
or bottom section.

More likely there was a variation in speed while cutting the ribbon, or the
light section was the first section of a new ribbon if two ribbons were
needed to get the six sections.  There are many other problems with
microtomy which could cause variations in section thickness.  My advice is
that when sectioning make sure the cutting speed is constant for all six
sections on the slide.

Good luck!

Joseph A. Saby, BA HT
Diagnostic Pathology, Pathology & Experimental Toxicology
Parke-Davis/Int. Pharm. Res. Div. Warner-Lambert
Ann Arbor, MI 48105
Phone: (734) 622-3631
Fax:     (734) 622-5718
e-mail:  Joseph.Saby@wl.com


-----Original Message-----
From: RBRIDDLE40@aol.com [mailto:RBRIDDLE40@aol.com]
Sent: Sunday, August 15, 1999 5:51 PM
To: histonet@pathology.swmed.edu
Subject: Need help


Hello Histo people,

We have Vashaw autostainer. We are having problem with uneven staining 
especially with our skins cases. We mount at least 6 or less frames on a 
slide. One of our pathologist is giving us a fit about the uneven staining. 
He may have a slide that is stain OK but one frame will be totally light
with 
very little staining. We have tried everything we know. Decrease the acid 
alcohol that we use, longer time in the hema.We dry them in an oven for 10 
min. and then on the stainer for ten min. They were lifting at one time. Any

help would be great.


Rita Riddle
Lexington Medical Center
West Columbia, SC